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Abstract:

A method and apparatus can separate and concentrate a selected component
from a multi-component material. The multi-component material may include
a whole sample such as adipose tissue, whole blood, or the like. The
apparatus generally includes a moveable piston positioned within a
separation container and a withdrawal tube that is operable to interact
with a distal end of the collection container past the piston. Material
can be withdrawn through the withdrawal tube.

Claims:

1. A method of separating a component from a selected material,
comprising: obtaining the selected material having multiple components;
providing a separation system including a tube having a top wall and a
bottom wall, a piston within the tube, an input port defined through the
top wall, an extraction port defined through the top wall, a hollow
member extending from the extraction port at least to the piston;
positioning the selected material into the separation system through the
input port with the top wall fixed to the tube and between the top wall
and the piston; centrifuging the separation system containing the
selected material; allowing the piston to move towards the top wall
during centrifugation; and extracting the component of the selected
material from between the piston and the bottom wall of the tube.

2. The method of claim 1: wherein positioning the selected material into
the separation system includes positioning the selected material
substantially only between the top wall and a first side of the piston.

3. The method of claim 2, wherein centrifuging the separation system and
allowing the piston to move towards the top wall includes: separating the
selected material into separate components; and allowing the piston to
move relative to a first of the components wherein at least a portion of
the first component is positioned between the bottom wall of the tube and
a second side of the piston.

4. The method of claim 3, wherein extracting the component includes
withdrawing at least a portion of the first component from between the
bottom wall of the tube and the second side of the piston through the
hollow member.

5. The method of claim 1, further comprising: providing the hollow member
substantially rigidly within the tube; providing a stop member extending
from the hollow member; and stopping the piston from moving towards the
top wall during centrifugation with the stop member.

6. The method of claim 1, further comprising: coupling a separate
injection port extender to the input port before positioning a selected
material into the separation system; and coupling a collection device
that collected the selected material to the injection port extender.

7. The method of claim 1, further comprising: coupling a collection
device to the input port of the separation system to position the
selected material into the separation system.

8. The method of claim 4, wherein withdrawing at least a portion of the
first component from between the bottom wall of the tube and the second
side of the piston through the hollow member further includes withdrawing
a portion of the first component through the piston.

9. A method of separating a component from a selected material,
comprising: obtaining the selected material with a collection device;
engaging the collection device to an input port of a separation system
having a container that includes a top wall and a bottom wall, a piston
within the container, and a withdrawal tube extending to the piston;
delivering the selected material through the input port of the separation
system with the collection device; removing the collection device from
the separation system; centrifuging the separation system containing the
selected material; and extracting the component of the selected material
from between the piston and the bottom wall of the container.

10. The method of claim 9, further comprising: allowing the piston to
move towards the top wall during centrifugation; and allowing the piston
to engage a stop member extending from the withdrawal tube to stop the
piston from moving towards the top wall.

11. The method of claim 9, wherein extracting the component includes
withdrawing at least a portion of the component from between the bottom
wall of the container and the piston through a withdrawal port in
communication with the withdrawal tube.

12. The method of claim 11, further comprising: withdrawing at least a
portion of the component through the piston in the withdrawal tube upon
coupling a withdrawal container to the withdrawal port.

13. The method of claim 9, wherein delivering the selected material
further includes positioning the selected material substantially only
between the top wall and a first side of the piston.

14. The method of claim 9, further comprising: allowing the piston to
move along the withdrawal tube during centrifugation until the piston
engages a stop member extending from the withdrawal tube.

15. A method separating a component from a selected material, comprising:
containing the selected material with a collection device; coupling the
collection device to an input port with a separation system having a
container that includes a top wall, a bottom wall and a piston movable on
a withdrawal tube within the container; delivering the selected material
through the input port of the separation system with the collection
device; moving the collection device from the separation system;
positioning the separation system in a centrifuge to apply a force to the
selected material in the separation system to allow a piston to move
towards the top wall of the container carrying centrifugation and
sequester the component between the piston and the bottom wall of the
container; and withdrawing the component from the separation system
through the withdrawal tube.

16. The method of claim 15, further comprising: Allowing the piston to
move towards a top wall during centrifugation until the piston engages a
stop member extending from the withdrawal tube.

17. The method of claim 15, wherein withdrawing the component further
includes withdrawing stromal cells from the separation system.

18. The method of claim 15, wherein sequestering the component from the
selected material further includes holding the piston at a selected
position relative to the withdrawal tube and holding the piston fixed
relative to the component of the selected material.

19. The method of claim 15, wherein withdrawing the component of the
selected material further includes coupling a withdrawal device to a
withdrawal port of the container and withdrawing the component through
the piston and the withdrawal tube.

20. The method of claim 15, further comprising: coupling an injection
port extender to the input port before coupling the collection device to
the input port of the separation system.

Description:

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is a division of U.S. patent application Ser. No.
13/285,436 filed on Oct. 31, 2011, which is a continuation of U.S. patent
application Ser. No. 11/744,093 filed on May 3, 2007, now U.S. Pat. No.
8,048,297 issued on Nov. 1, 2011, which is (a.) a continuation-in-part of
U.S. patent application Ser. No. 11/210,005 filed on Aug. 23, 2005, now
U.S. Pat. No. 7,771,590 issued on Aug. 10, 2010; and (b.) also claims the
benefit of U.S. Provisional Application No. 60/900,758, filed on Feb. 9,
2007. The disclosures of the above applications are incorporated herein
by reference.

FIELD

[0002] The present teachings relate generally to collection of selected
biological materials, in particularly to a method and apparatus for
separating and collecting a selected biological component.

BACKGROUND

[0003] Various biological materials, such as whole blood, adipose tissue
and the like, are formed of a plurality of components or fractions. These
various fractions can be collected and separated from an anatomy, such as
a human anatomy, using various techniques. Nevertheless, generally known
techniques may require a plurality of steps and a large volume of
biological materials to obtain a selected biological component.

[0004] For example, collecting a selected component of whole blood or
adipose tissue requires collecting a large sample of whole blood or whole
adipose tissue and performing several steps to obtain a selected fraction
of the whole sample. It may be desirable to obtain a selected volume for
a procedure where time and sample quantity are minimal. Therefore, it may
be desirable to provide a method and apparatus to obtain a selected
volume of a fraction of a biological material in a short period of time
from a selected volume.

SUMMARY

[0005] A method and apparatus is provided for obtaining a selected
fraction or component of a biological material for a use. The apparatus
can generally include a container and a solid or porous piston. A
withdrawal tube can be permanently or selectively interconnected with the
piston to withdraw a selected fraction of a whole material. Generally,
the withdrawal tube can pass through a selected portion of the piston,
such as a distal end of the piston to obtain a material that is
positioned near a distal portion of the container.

[0006] According to various embodiments a system to separate a component
from a selected material is disclosed. The system can include a
separation container operable to contain the selected material having a
top and a bottom and a top wall at a proximal end of the separation
container that closes the top of the separation container. A piston can
be positioned in the separation container. An injection port can extend
through the top wall. In addition, a conduit can be positioned in the
separation container operable to remove the selected material from a
distal end near the bottom of the separation container past the piston.

[0007] According to various embodiments a system to separate a component
from a selected material is disclosed. The system can include a container
having a side wall, bottom wall, and a top wall and defining an interior
volume. An input port can extend from the top wall and define a first
passage through the top wall to the interior volume. An extraction port
can extending from the top wall. A piston can move within the interior
volume of the container. In addition, a conduit extending from the
extraction port can include a tube extending from the top wall and a
passage through the piston.

[0008] According to various embodiments, a method of separating a
component from a selected material is disclosed. The method can include
obtaining the selected material having multiple components and providing
a separation system including a tube having a top wall, a piston within
the tube, an input port defined through the top wall, an extraction port
defined through the top wall, a hollow member extending from the
extraction port at least to the piston. The selected material can be
positioned in the separation system through the input port with the top
wall connected to the tube and between the top wall and the piston. The
separation system can be centrifuged while containing the selected
material and the piston can move towards the top wall during
centrifugation. The component of the selected material can be extracted
from past the piston.

[0009] Further areas of applicability of the present teachings will become
apparent from the detailed description provided hereinafter. It should be
understood that the detailed description and various embodiments are
intended for purposes of illustration only and are not intended to limit
the scope of the teachings.

BRIEF DESCRIPTION OF THE DRAWINGS

[0010] The present teachings will become more fully understood from the
detailed description and the accompanying drawings, wherein:

[0011] FIG. 1 is a kit of an apparatus according to various embodiments;

[0012] FIG. 2 is an environmental view of a separating device according to
the various embodiments;

[0013] FIG. 3 illustrates the separating device being filled according to
various embodiments;

[0014]FIG. 4 is an environmental view of a filled separating device
according to various embodiments;

[0015] FIG. 5 is an environmental view of a separating device at a
centrifuge according to various embodiments;

[0016]FIG. 6 is an environmental view of a separating device after being
centrifuged;

[0017]FIG. 6A is a schematic view of a separating device after being
centrifuged;

[0018] FIG. 7 is an environmental view of material being withdrawn from
the separating device according to various embodiments;

[0019]FIG. 7A is a schematic view of the piston in the container while
material is being withdrawn from the separating device according to
various embodiments;

[0020]FIG. 8 illustrates the environmental view after a selected
component has been withdrawn from the separating device;

[0021] FIG. 9 is an exploded perspective view of a separation device
according to various embodiments;

[0022] FIG. 10 is an assembled view of a separation device according to
various embodiments; and

[0023] FIG. 11 is a detail view of a syringe interacting with a separation
device according to various embodiments.

DETAILED DESCRIPTION OF VARIOUS EMBODIMENTS

[0024] The following description of the various embodiments is merely
exemplary in nature and is in no way intended to limit the teachings, its
application, or uses. Although the following teachings relate to adipose
tissue, it will be understood that the teachings may apply to any
appropriate multi-component material, whether biological or not. It will
be further understood that a component can be any appropriate portion of
a whole, whether differing in density, specific gravity, buoyancy,
structure, etc. The component is a portion that can be separated from the
whole.

[0025] With reference to FIG. 1, a kit 20 can be provided to allow for
collection, separation, and application of a selected biological material
or component. The kit 20 can be understood to include any appropriate
devices or materials, and the following devices are merely exemplary. The
kit 20 can include a separation device 30 that can be used to separate a
selected material, such as an adipose tissue sample, a whole blood
sample, or the like. It will be understood that the separation device 30
can be disposable, reusable, or combinations thereof. For example, the
separation device 30 can include a container 32 that may be reusable
while a separation piston 34 is not. Further, the kit 20 can include a
collection device such as a syringe 36, an application device such as a
syringe 38, and a mixing material that may be included in a syringe 40.
The mixing material may be any appropriate material such as an
anti-clotting agent, a clotting agent, an antibiotic, an enzyme, a
buffer, a growth factor or factors, or the like. It will be understood
that the kit 20 may also include any other appropriate materials such as
bandages, tourniquets, sterilization materials or the like. It will be
further understood that the kit 20 may be provided sterilized, prepared
for sterilization, or any appropriate combination thereof.

[0026] The various syringes 36, 38, 40, may be any generally known
syringe. The syringe 36 may also be interconnectable with a needle or
cannula 42 that can interconnect with a luer fitting 44 of the syringe
36. The syringe 36 can generally include a container 46 and a plunger 48.
This can allow the syringe 36 to withdraw a selected sample, such as an
adipose tissue sample from an anatomy, such as a human anatomy, for
various purposes. The application syringe 38 can also include a container
50 and a plunger 52. The application syringe 38 can be any appropriate
syringe and can be of a size to interconnect with the selected portion of
the separation device 30, such as discussed herein. Further, the mixing
syringe 40 can also include a container 54 and a plunger 56. The mixing
syringe 40 can include any appropriate material, such as those described
above. The mixing material provided in the mixing syringe 40. The mixing
material can be added to the container 32 at any appropriate time for
interaction with the selected material that can also be positioned in the
separation container 32.

[0027] The separation device 30 includes the container or tube 32 that can
include various features. For example, container 32 can be any
appropriate size such as 20 ml, 40 ml, 60 ml, any combination thereof,
fraction thereof, or any appropriate size. The collection container 32
includes a side wall 60 that can assist in containing the material
positioned in the container 32. The tube 32 may also include demarcations
62 that indicate a selected volume.

[0028] The sidewall 60 may or may not be flexible under a selected force.
For example, the separation device 30 can be positioned in a centrifuge
or similar device to apply an increased force of gravity to the material
positioned in the tube 32. If the tube 32 is formed of a selected
material, the sidewall 60 may flex under the high force of gravity to
cause an increased diameter of the tube 32 under the higher force of
gravity. Alternatively, the sidewall 60 of the container 32 may be formed
of a substantially rigid material that will not flex under a high force
of gravity.

[0029] The tube 32 further includes a top or proximal portion that defines
a cap engaging region 64. The cap engaging region 64 can include a thread
or partial threads 66 that can interconnect with a cap 68. The cap 68 can
include an internal thread that can thread onto the thread 66 of the top
portion 64 to fix the cap 68 relative to the tube 32. Therefore, the cap
68 can be removed from the tube 32, but it will be understood that the
cap 68 can also be formed as an integral or single portion of the tube
32. It will be understood that the separating device 30 can be provided
as a modular system or can be formed as an integral or unitary member.

[0030] Extending through the cap 68 can be a collection or application
port 72. The port 72 can include a luer locking portion 74, or any other
appropriate interconnection portion. The port 72 can also include or be
connected to a cap 71. The port 74 can extend through the cap 68 to a
withdrawal tube 76. The withdrawal tube 76 may be formed as a single
piece with the port 72 or can be interconnectable with the port 72.
Further, the withdrawal tube 76 can extend through the piston 34 through
a central channel 78 defined through the piston 34. The withdrawal tube
76 can define a conduit, such as an extraction conduit. One skilled in
the art will understand that a separate tube or cannula can be passed
relative to the piston 34 for withdrawal of a material or component of
the sample. Thus, the withdrawal tube 76 need not be maintained in the
tube 32 for an entire procedure.

[0031] The withdrawal tube 76 can, but is not required to, define a piston
stop or stop member 80. The stop 80 can act as a stop member for the
piston 34 so that the piston 34 is able to move only a selected distance
along the withdrawal tube 76. The stop 80 can also be formed by any
appropriate portion, such as the sidewall 60. The stop 80 is provided to
assist in limiting a movement of the piston 34. Therefore, it will be
understood that the withdrawal tube 76 may also act as a rod on which the
piston 34 is able to move.

[0032] The piston 34 can include any appropriate geometry such as a
geometry that substantially mates with the tube 32, particularly a distal
end 82 of the tube 32. The distal end of the tube 32 can be flat,
conical, tapered, etc. It will be understood, however, that the piston 34
can also include any other appropriate geometry to interact with the tube
32. Further, the piston 34 can include a contacting or central region 84
that includes an outer dimension, such as a circumference or diameter
that is generally equivalent to an inner diameter or circumference of the
tube 32. Therefore, the piston 34 can contact or engage the sidewall 60
of the tube 32 at a selected time.

[0033] The piston 34 can also be formed in any appropriate configuration
or of any appropriate material. For example, in addition to the selected
geometry of the piston 34, the piston can be porous, non-porous, or
include regions of each. For example, the piston 34 can be formed of a
porous material such as a screen, a filter, a mesh, or the like. The
piston 34, including a porous region, can allow a selected material to
pass through and not allow other non-selected materials to pass. The
piston 34, therefore, can selectively separate materials or components of
a sample.

[0034] The middle or tube engaging portion 84 of the piston 34 can include
the dimension that is substantially similar to an unchanged or unforced
dimension of the wall 60 of the tube 32. For example, it may be formed so
that there is substantially little space or a sliding engagement between
the tube engaging portion 84 of the piston 34 and the tube 32. However,
under a selected force, such as a centrifugal force, the wall 60 of the
tube 32 can be compressed axially and be forced outward thereby
increasing a dimension, such as a diameter, of the tube 32. The
increasing of the diameter of the tube 32 relative to the piston 34 can
allow for a freer movement or non-engagement of the tube 32 with the
piston 34. In this way, the piston 34 can move relative to the tube 32 or
materials can move between the piston 34 and the tube 32.

[0035] For example, as discussed herein, the piston 34 may move relative
to the tube 32 when the tube is compressed, thus increasing the tube's 32
diameter. The piston 34 can move relative to the withdrawal tube 76,
which can allow the piston 34 to move a selected distance relative to the
tube 32 or the cap 68. The stop 80, which is provided on the withdrawal
tube 76, can assist in selectively stopping the piston 34 relative to the
rod 76. This can define a maximum motion of the piston 34 relative to the
withdrawal tube 76.

[0036] A selected material, such as a biological material, can be
positioned in the tube 32 and the tube 32 can be positioned in a
centrifuge with the piston 34. During the centrifugal motion, the tube 32
can compress, thereby increasing its diameter relative to the piston 34.
The compression can allow the piston 34 to more easily move relative to
the withdrawal tube 76 and the container tube 32. Therefore, the piston
34 can assist in separating a selected material positioned in the
container tube 32. Once the centrifugal force is removed or reduced, the
axial compression of the container tube 32 can be reduced to thereby
return it substantially to its original dimensions. As discussed above,
its original dimensions can be substantially similar to those of the
piston 34, particularly the tube engaging portion 84, which can hold the
piston 34 in a selected position relative to the tube 32. This can assist
in maintaining a separation of the material positioned in the tube 32, as
discussed herein.

[0037] It will be understood that the separation system 30 can be used
with any appropriate process or various selected biological materials or
multi-component materials. Nevertheless, the separation system 30 can be
used to separate a selected biological material such as stromal cells,
mesenchymal stem cells, blood components, adipose components or other
appropriate biological or multi-component materials. Thus, it will be
understood that the following method is merely exemplary in nature and
not intended to limit the teaching herein.

[0038] With additional reference to FIG. 2, a patient 90 can be selected.
The patient 90 can include an appropriate anatomy and the collection
device 36 can be used to collect a selected portion of biological
material. For example, the collection device 36 can engage a portion of
the patient 90 to withdraw a selected volume of adipose tissue. The
adipose tissue can be selected from any appropriate portion of the
anatomy, such as from the abdominal region. In addition, various other
components may be withdrawn into the collection tube 36, such as whole
blood, stem cells, and the like. Further, the collection device 36 can be
a plurality of collection devices that each collect different components,
such as one to collect adipose tissue, one to collect whole blood, and
others to collect other selected biological materials.

[0039] Once the selected biological material is withdrawn into the
collection device 36, the biological material 92 can be placed into the
tube 32. Once the tube 32 has been filled an appropriate amount with the
biological material 92, the piston 34, the rod 76, and the cap 68 can be
interconnected with the tube 32.

[0040] With additional reference to FIG. 4, the assembled separation
device 30 can be pre-treated prior to various other processing steps. For
example, selected components, including enzymes, chemicals, antibiotics,
growth factors, and the like, can be added to the container tube 32.
Further, the selected material, which can include adipose tissue, can be
sonicated or treated with a sonic radiation prior to further processing
steps. In addition, or alternatively to sonication, various other
agitating methods or devices can be used to mix or agitate the material.
For example, a mixing bead, beads, ball, or the like can be placed in the
container 32. The container 32 can then be moved with the beads inside to
agitate and mix the material. In addition, various rigid arms or
extensions can be positioned in the container 32 to assist in agitating
or mixing the material.

[0041] The sonication of the adipose tissue can perform various steps. For
example, the sonication of the adipose tissue can remove or release
stromal cells from the adipose tissue cells. It will be understood that
sonication of the adipose tissue can be performed at any appropriate
time. For example, the sonication of the adipose tissue can be performed
once it has been collected into the collection device 36 and prior to
being positioned in the tube 32 or after it has been positioned in the
tube 32. Further, all of the selected materials, which may include whole
blood, various components of whole blood, or the like, can be also added
to the tube 32.

[0042] With reference to FIG. 5, once the separation system 30 has been
optionally pre-processed, such as with agitation and/or sonication,
various chemicals, various biologically active materials (e.g. enzymes),
it can be positioned in an appropriate separation device, such as a
centrifuge 94. The centrifuge 94 can be operated according to any
appropriate technique to perform a high gravity separation of the
material positioned in the separation device 30. Nevertheless, the
centrifuge device can be spun at any appropriate rotation per minute
(RPM) such as about 2000 to about 4030 RPMs. This can form a force of
gravity on the separation device 30 and the various materials positioned
therein of about 740 G's to about 3000 G's. Further, the centrifugation
step with the centrifuge device 94 can be performed for any appropriate
amount of time. For example, the separation device 30 can be spun at the
selected RPMs for about 5 to about 15 minutes. It will be understood that
one skilled in the art can determine an appropriate RPM and time setting
which can be used to separate selected materials positioned in the
separation device 30. Further, the separation of different materials may
require different RPMs and different separation times.

[0043] As discussed above, the piston 34 can be positioned in the tube 32
to assist in separating the materials positioned in the container tube
32. The piston 34 can be formed of any appropriate materials and
according to any appropriate physical characteristics. For example, the
piston 34 can be formed of a material or combination of materials that
can achieve a selected density. The piston 34 can assist in separating,
such as physically separating, selected components of the biological
material 92 positioned in the separation device 30. For example, the
piston 34 can include a density that is about 1.00 grams per milliliter
to about 1.10 grams per milliliter, such as less than about 1.06 grams
per cc or 1.06 grams per milliliter. The selected density of the piston
34 can assist in separating denser components or components with a higher
specific gravity than the piston 34. For example, stromal cells include a
specific gravity that is greater than other components of the biological
material 92 positioned in the tube 32 and also greater than that of the
piston 34. The piston 34, however, can include any appropriate density.

[0044] As discussed above, when the separation device 30 is positioned in
the centrifuge 94 the centrifuge 94 can be spun. The forces produced by
the centrifuge 94 can compress the container tube 32, which can increase
its diameter thus allowing the piston 34 to move relative to the
container 32. The various components of the biological material 92
positioned in the separation tube 32 can be physically separated by the
piston 34 as it moves relative to the separation tube 32. This can assist
in moving at least one of the piston 34 or a portion of the biological
material 92. Though the biological material can originally be positioned
on top of the piston 34, the forces and/or flexing of the sidewall 60 can
allow at least a component of the material to move past the piston 34. It
will be understood, however, that the sidewall 60 may not flex and that
the material is simply forced past the piston 34 between the piston 34
and the sidewall 60. Thus, it will be understood that the material can
move past the piston 34 to the distal end 82 to container 32 according to
any appropriate method such as flexing the sidewall 60, moving between a
space between the piston 34 and the sidewall 60, or any other appropriate
method.

[0045] With additional reference to FIG. 6, the biological material 92 can
be separated into a plurality of components that are contained within the
separation container 32. For example, a first component 92a can be
positioned between the piston 34, such as a distal end of the piston 34a
and the distal end of the separation container 82. The first biological
component 92a can be any appropriate material, including stromal cells,
mesenchymal stem cells or the like. If the biological material 92
positioned within the separation tube 32 includes adipose tissue, then
various other components can include a plasma and plasma protein
component 92b and a fat and oil component 92c. It will be understood, as
illustrated in FIG. 6, that the fat and oil component 92c is generally
formed near a proximal end of the tube 32 while the denser stromal cells
are formed as a cell button near the distal and 82. Further, it will be
understood that various materials, including plasma and plasma proteins,
may also include a density that is higher than that of the piston 34 and
thus may also be formed or moved towards the distal end 82 of the
separation tube 32. Nevertheless, the first component 92a can include a
high concentration of the high density materials that is of a selected
material to be separated using the separation device 30, because of the
piston 34 and the stop 80.

[0046] Further, because the various materials, such as plasma or plasma
proteins, can include a density that is similar to that of the first
component 92a, which can include the stromal cells, the stop 80 can
extend from the withdrawal tube 76 to ensure a low concentration or low
volume of the plasma, plasma proteins, or the materials that may include
a density that is greater than that of the piston 34. Although it may be
selected to include a selected volume of the plasma or plasma proteins
near the distal end 82 of the separation tube 32, such as for withdrawal
of the selected cells, such as stromal cells, it may be selected to keep
the concentration at a selected amount. Therefore, the stop 80 or other
stop or limiting portion (e.g. a lip or edge in the container 32) can
assist in achieving the selected volume and concentration of the first
component 92a to be separated by the separation device 30 as the piston
34 moves towards the stop 80, as illustrated in FIGS. 6 and 6A, where the
piston 34 is illustrated to have moved away from the distal end 82 of the
container 32.

[0047] With additional reference to FIG. 7, the withdrawal device 38 can
be interconnected with the withdrawal port 72 which interconnects the
withdrawal device 38 with the withdrawal tube 76. As discussed above, the
withdrawal tube 76 can pass through the piston 34. Because the withdrawal
tube 76 can be fixed relative to the cap 78, the withdrawal tube 76 may
not move during the centrifugation process. This allows the piston 34 to
move relative to the separation tube 32 while the withdrawal tube 76
maintains its position, as illustrated in FIGS. 6, 6A, and 7. The
withdrawal tube 76 can include a portion positioned generally near the
distal portion 82 of the separation tube 32. Therefore, the withdrawal
port 72 can be interconnected or operable to remove a material that is
positioned near the distal end 82 of the separation tube 32. Though the
piston 34 can move proximally and allow for separation of a volume near
the distal end 82 of the separation tube 32, the withdrawal tube 76 is
still positioned near the distal end 82 of the separation tube 32.
Therefore, the collection device 38 can be interconnected with the
withdrawal port 72 and used to withdraw the volume of material that is
positioned near the distal end of the tube 82, as illustrated in FIGS. 6,
6A, and 7. Thus, the separated material, which can include stromal cells
or other appropriate biological components, can be withdrawn after being
separated and concentrated with the separation system 30. Other various
components, such as the components 92b and 92c of the biological material
92 can be retained in the tube 32.

[0048] As the collection device 38 withdraws material from the separation
tube 32, the piston 34 can be moved generally in the direction of the
arrow A, as illustrated in FIGS. 7 and 7A, away from the stop 80. This
can allow for a displacement of the volume being removed into the
collection tube 38 as the piston 34 moves in the direction of arrow A
towards the distal end 82 of the separation tube 32. Further, this
movement of the piston 34 can assist in withdrawing the material from the
distal end 82 of the separation tube 32.

[0049] With reference to FIGS. 7A and 8, the piston 34 can remain or,
again, move to substantially fill the internal volume of the distal
portion 82 of the separation tube 32 as it moves toward the distal end 82
as the component is withdrawn. Therefore, the piston 34 can also assist
in withdrawing the material from the separation tube 32. Since the piston
34 can substantially fill the volume of the material 92a being withdrawn
from the separation tube 32, it can help insure that substantially all of
the volume of the material 92a is withdrawn from the separation container
32.

[0050] Therefore, the separation device 30 can assist in separating,
concentrating, and collecting a selected biological component of the
biological material 92. It will be understood that while collecting
stromal cells from a sonicated adipose tissue is described that the
separation, concentration, and collection of any selected biological
component may be performed. One skilled in the art will understand that
the separation device 30 can be used with any appropriate biological
material that can be positioned in the separation tube 32.

[0051] The separation device 30 can be used to separate and concentrate a
selected volume of material from a substantially small volume of the
whole biological material 92. Because the separation system 30 includes
the various components, including the withdrawal tube 76 that extends
substantially the length of the separation container 32, and the piston
34, the biological material 92 can be effectively separated and
concentrated into various components. The denser component 92a can be
easily withdrawn from the separation tube 32 without interference of the
other components of the biological material 92.

[0052] The withdrawn material, which may include the stromal cells, can
then be used for various purposes. The withdrawn material can include the
selected biological component, such as stromal cells, mesenchymal stem
cells, or other stem cells. The stromal cells that are collected from the
selected biological material, such as adipose tissue, can be applied to
various portions of the anatomy to assist in healing, growth,
regeneration, and the like. For example, during an orthopedic procedure,
an implant may be positioned relative to a bony structure. The stromal
cells or other components can be applied near the site of the
implantation, to the implant before implantation, to an area of removed
bone, or the like, to assist in regeneration of growth of the bone. The
stem cells, such as the stromal or mesenchymal cells, can assist in
healing and growth of the resected bone. Therefore, the separated and
concentrated biological component, which can include the stromal cells or
other appropriate biological components, can be applied to assist in
regeneration, speed healing after a procedure, or other appropriate
applications. Briefly, the undifferentiated cells can differentiate after
implantation or placement in a selected portion of the anatomy.
Alternatively, the cells can release factors that direct the activity of
other cells to assist in regeneration, speed healing, or other
appropriate applications.

[0053] With reference to FIGS. 9 and 10, the kit 20 can include a
separation device 100 that is similar to the separation device 30. While
the separation device 100 differs from the separation device 30 in
various aspects those identical portions will be referenced with
identical reference numerals. Briefly, the separation device 100 can
include the separation container 32 or tube. Further, the separation
device 100 can include the piston 34. The piston 34 can be positioned
within the tube 32 of the separation device 100. The separation device
100 can also include the cap or top wall 68. According to various
embodiments, the top wall 68 can be substantially fixed to a proximal end
102 of the tube 32. As discussed above, the top wall 68 can also
threadably engage a cap engaging region 64 of the tube 32. An adhesive
can be used to fix the cap or top wall 68 to the proximal end 102 of the
tube 32 or the two can be formed as a single member.

[0054] The separation device 100 can differ from the separation device 30
according to various features. For example, the separation device 100 can
include an injection port or second port 104. The injection port 104 can
extend between an outlet end 106 and an inlet end 108. The inlet end 108
can also include a connection portion, such as a quarter turn or luer
connection that can interconnect with an injection port extender 110. The
injection port extender 110 can include a top or injection end 112. A cap
114 can be positioned over the top 112 of the extension 110. The top 112
can include a connection portion, such as a luer lock or other connection
portion to connect with the cap 114 or an injection syringe, as discussed
further herein.

[0055] The separation device 100 can also include a second injection port
cap 116. The second injection port cap 116 can be tethered to the top
wall 68 with a tether 118. The second injection port cap 116 can also
include a sterile contact or holding member 120 that can be removed after
use. The second injection port cap 116 can include a luer connection or
fixation port to connect to the injection port 104 at the top or
connection portion 108.

[0056] The injection port 104 allows the material to be injected through
the top wall 68 into the tube 32. The top wall 68 can, therefore, be
fixed to the proximal end 102 of the tube 32 while the material is being
injected or delivered to the tube 32. This can allow the multi-component
material 92 to be delivered into the tube 32 in an efficient manner and
can also maintain the position of the piston 34 near the distal end 82 of
the tube 32. Also, any appropriate mixing material can be added at any
appropriate time from the syringe 40 or other source. According to
various embodiments, the top wall or cap 68 can be removed a small amount
and the material 92 can be delivered through the top end or proximal end
102 of the tube 32. Providing the injection port 104, however, can
provide a mechanism and port to inject the material into the injection
tube without removing the cap 68 from the tube 32.

[0057] With additional reference to FIG. 11, the collection device or
syringe 36 can be interconnected with the extension 110 that is
interconnected with the injection port 104. The collection syringe 36, as
discussed above, can be used to collect the multi-component fluid 92. The
multi-component fluid 92 can be injected into the tube 32 of the
separation device 100. The separation device 100 can include the top wall
68 substantially fixed to the tube 32. The extraction port 72 can also be
positioned relative to the cap 68 and be interconnected with the conduit
76.

[0058] The extension 110 can allow the collection syringe 36 to be
interconnected with the injection port 104 in a manner that allows access
without interference of the extraction port 72. The extension 110, as
discussed above, can include the luer connection near the top end 112 of
the extension 110 to interconnect with the collection syringe 36.
Therefore, the syringe 36 can be efficiently connected to the extension
110 which is connected to the injection port 104.

[0059] Once the material is injected into the tube 32 through the
injection port 104, the extension 110 can be removed from the injection
port 104. After the extension 110 is removed from the injection port 104,
the second injection port cap 116 can be interconnected with the
injection port 104. The sterile holder 120 on the second injection port
cap 116 can be used to effectively maintain sterility between the second
injection port cap 116 and the injection port 104. The second injection
port cap 116 can be positioned over the injection port 104 during the
centrifugation process and the extraction process from the tube 32.

[0060] The separation device 100 can be used in a manner substantially
identical to the separation device 30, discussed above. It will be
understood that the extension 110 is not required, and can be provided
according to various embodiments or when selected by a user. Further, the
separation device 100 can be included in the kit 20, either with the
separation device 30 or as an alternative thereto. Therefore, one skilled
in the art will understand, the separation device 100 can be included
with the kit 20 and used as the separation device 30 discussed above. In
addition the separation devices 30, 100 and the kit 20 can be used in
various procedures, such as wound healing, including stromal cells from
adipose tissue and other blood components, as taught in U.S. Provisional
Application No. 60/900,758, filed on Feb. 9, 2007, incorporated herein by
reference.

[0061] The teachings are merely exemplary in nature and, thus, variations
that do not depart from the gist of the teachings are intended to be
within the scope of the teachings. Such variations are not to be regarded
as a departure from the spirit and scope of the teachings.